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1.
Dev Biol ; 516: 158-166, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-39173813

RESUMO

The function of spargel/dPGC-1 in Drosophila oogenesis has been unequivocally established. Here, we sought to assess whether Spargel protein or RNA is essential for developmentally competent eggs. The trans-heterozygotic combination of two spargel mutant alleles allowed us to decrease Spargel expression to very low levels. Using this model, we now demonstrated the requirement for Spargel in eggshell patterning and embryonic development, which led us to establish that spargel is a maternal effect gene. Further examination of Spargel's potential mechanism of action in eggshell biogenesis revealed that low levels of Spargel in the adult ovary cause diminished Cyclin E activity, resulting in reduced chorion gene amplification levels, leading to eggshell biogenesis defects. Thus, another novel role for spargel/dPGC-1 is exposed whereby, through Cyclin E activity, this conserved transcriptional coactivator regulates the chorion gene amplification process.


Assuntos
Córion , Ciclina E , Proteínas de Drosophila , Desenvolvimento Embrionário , Amplificação de Genes , Fator B de Elongação Transcricional Positiva , Animais , Feminino , Córion/metabolismo , Córion/embriologia , Ciclina E/metabolismo , Ciclina E/genética , Drosophila melanogaster/embriologia , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Desenvolvimento Embrionário/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Mutação , Oogênese/genética , Fator B de Elongação Transcricional Positiva/genética , Fator B de Elongação Transcricional Positiva/metabolismo
2.
Genesis ; 62(2): e23592, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38587195

RESUMO

Mesenchymal stem cells (MSCs) derived from fetal membranes (FMs) have the potential to exhibit immunosuppression, improve blood flow, and increase capillary density during transplantation. In the field of medicine, opening up new avenues for disease treatment. Chicken embryo chorioallantoic membrane (CAM), as an important component of avian species FM structure, has become a stable tissue engineering material in vivo angiogenesis, drug delivery, and toxicology studies. Although it has been confirmed that chorionic mesenchymal stem cells (Ch-MSCs) can be isolated from the outer chorionic layer of FM, little is known about the biological characteristics of MSCs derived from chorionic mesodermal matrix of chicken embryos. Therefore, we evaluated the characteristics of MSCs isolated from chorionic tissues of chicken embryos, including cell proliferation ability, stem cell surface antigen, genetic stability, and in vitro differentiation potential. Ch-MSCs exhibited a broad spindle shaped appearance and could stably maintain diploid karyotype proliferation to passage 15 in vitro. Spindle cells were positive for multifunctional markers of MSCs (CD29, CD44, CD73, CD90, CD105, CD166, OCT4, and NANOG), while hematopoietic cell surface marker CD34, panleukocyte marker CD45, and epithelial cell marker CK19 were negative. In addition, chicken Ch-MSC was induced to differentiate into four types of mesodermal cells in vitro, including osteoblasts, chondrocytes, adipocytes, and myoblasts. Therefore, the differentiation potential of chicken Ch-MSC in vitro may have great potential in tissue engineering. In conclusion, chicken Ch-MSCs may be an excellent model cell for stem cell regenerative medicine and chorionic tissue engineering.


Assuntos
Galinhas , Células-Tronco Mesenquimais , Animais , Embrião de Galinha , Membrana Corioalantoide , Diferenciação Celular/fisiologia , Células Cultivadas
3.
J Biol Chem ; 299(4): 104600, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36906145

RESUMO

Teleost oocytes are surrounded by a structure called chorion or egg envelopes, which is composed of zona pellucida (ZP) proteins. As a result of the gene duplication in teleost, the expression site of the zp genes, coding the major component protein of egg envelopes, changed from the ovary to the maternal liver. In Euteleostei, there are three liver-expressed zp genes, named choriogenin (chg) h, chg hm, and chg l, and the composition of the egg envelope is mostly made up of these Chgs. In addition, ovary-expressed zp genes are also conserved in the medaka genomes, and their proteins have also been found to be minor components of the egg envelopes. However, the specific role of liver-expressed versus ovary-expressed zp genes was unclear. In the present study, we showed that ovary-synthesized ZP proteins first form the base layer of the egg envelope and then Chgs polymerize inwardly to thicken the egg envelope. To analyze the effects of dysfunction of the chg gene, we generated some chg knockout medaka. All knockout females failed to produce normally fertilized eggs by the natural spawning. The egg envelopes lacking Chgs were significantly thinner, but layers formed by ZP proteins synthesized in the ovary were found in the thin egg envelope of knockout as well as wildtype eggs. These results suggest that the ovary-expressed zp gene is well conserved in all teleosts, including those species in which liver-derived ZP proteins are the major component, because it is essential for the initiation of egg envelope formation.


Assuntos
Proteínas de Peixes , Fígado , Oryzias , Ovário , Glicoproteínas da Zona Pelúcida , Animais , Feminino , Sequência de Aminoácidos , Fígado/metabolismo , Oryzias/genética , Oryzias/metabolismo , Ovário/anatomia & histologia , Ovário/metabolismo , Zona Pelúcida/química , Zona Pelúcida/metabolismo , Glicoproteínas da Zona Pelúcida/genética , Glicoproteínas da Zona Pelúcida/metabolismo , Técnicas de Inativação de Genes , Expressão Gênica , Óvulo/citologia , Óvulo/metabolismo
4.
Artigo em Inglês | MEDLINE | ID: mdl-39263983

RESUMO

False clownfish (Amphiprion ocellaris) employ a hatching strategy regulated by environmental cues, wherein parents provide water flow to encourage embryos to hatch after sunset on the hatching day. Despite previous studies demonstrating the necessity of complete darkness and water agitation for hatching, the regulatory mechanisms underlying these environmental cues remain elusive. This study aimed to investigate how darkness and water agitation affect the secretion of hatching enzymes and the hatching movements of embryos in false clownfish. Assessment of chorion digestion and live imaging of Ca2+ in hatching glands using GCaMP6s, a Ca2+ indicator, revealed that darkness stimulation triggers the secretion of hatching enzymes by increasing Ca2+ levels in hatching gland cells. On the other hand, water agitation primarily stimulated hatching movements in embryos, which led to the rupture of their egg envelopes. These results suggest that changes in light environments following sunset induce embryos to secrete hatching enzymes and that water agitation provided by parents stimulates hatching movements. These responses to environmental cues, light and water agitation, contribute to the rapid and synchronous hatching in false clownfish.

5.
Gen Comp Endocrinol ; 349: 114453, 2024 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-38281702

RESUMO

The prss59.1 gene was identified as one of 11 genes that were highly upregulated during the induction of ovulation in zebrafish by using an in vivo ovulation assay. Previously, we conducted biochemical characterization of Prss59.1 and revealed it to be a trypsin-like proteolytic enzyme. In this study, we established a prss59.1 gene knockout strain using the CRISPR/Cas9 system. Phenotypic analysis of prss59.1 knockout fish showed that prss59.1 is associated with chorion elevation, a prominent event in egg activation during fertilization. The chorions of heterozygous and homozygous prss59.1 mutant zebrafish were smaller than those of the wild type. The results suggested that Prss59.1 is necessary for chorion expansion. The homozygous prss59.1 mutant strain, with a small chorion, showed an extremely low survival rate. Fiber-supported knob-like structures (KS) on the chorion showed an abnormal structure in prss59.1 mutants. Prss59.1 was detected in the KS on the chorion. The pores on the chorion were smaller in the prss59.1 mutants than in the wild type. Transmission electron microscopy (TEM) observations of the cross sections of the chorions showed abnormalities in the chorion structure in prss59.1 mutants. These results demonstrated that Prss59.1 is involved in chorion elevation and in proper formation of the chorion, which is necessary for embryo development.


Assuntos
Fertilização , Peixe-Zebra , Animais , Feminino , Peixe-Zebra/fisiologia , Homozigoto , Córion/química , Córion/fisiologia
6.
Arch Gynecol Obstet ; 309(5): 1919-1923, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-37184578

RESUMO

PURPOSE: The fetal membranes are essential for the maintenance of pregnancy, and their integrity until parturition is critical for both fetal and maternal health. Preterm premature rupture of the membranes (pPROM) is known to be an indicator of preterm birth, but the underlying architectural and mechanical changes that lead to fetal membrane failure are not yet fully understood. The aim of this study was to gain new insights into the anatomy of the fetal membrane and to establish a tissue processing and staining protocol suitable for future prospective cohort studies. METHODS: In this proof of principle study, we collected fetal membranes from women undergoing vaginal delivery or cesarean section. Small membrane sections were then fixed, stained for nucleic acids, actin, and collagen using fluorescent probes, and subsequently imaged in three dimensions using a spinning disk confocal microscope. RESULTS: Four fetal membranes of different types were successfully processed and imaged after establishing a suitable protocol. Cellular and nuclear outlines are clearly visible in all cases, especially in the uppermost membrane layer. Focal membrane (micro) fractures could be identified in several samples. CONCLUSION: The presented method proves to be well suited to determine whether and how the occurrence of membrane (micro) fractures and cellular jamming correlate with the timing of membrane rupture and the mode of delivery. In future measurements, this method could be combined with mechanical probing techniques to compare optical and mechanical sample information.


Assuntos
Ruptura Prematura de Membranas Fetais , Nascimento Prematuro , Feminino , Recém-Nascido , Gravidez , Humanos , Cesárea , Estudos Prospectivos , Córion , Membranas Extraembrionárias , Microscopia Confocal
7.
J Wound Care ; 33(Sup7): S4-S14, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38973638

RESUMO

OBJECTIVE: Diabetic foot ulcers (DFUs) continue to challenge wound care practitioners. This prospective, multicentre, randomised controlled trial (RCT) evaluated the effectiveness of a dehydrated Amnion Chorion Membrane (dACM) (Organogenesis Inc., US) versus standard of care (SoC) alone in complex DFUs in a challenging patient population. METHOD: Subjects with a DFU extending into dermis, subcutaneous tissue, tendon, capsule, bone or joint were enrolled in a 12-week trial. They were allocated equally to two treatment groups: dACM (plus SoC); or SoC alone. The primary endpoint was frequency of wound closure determined by a Cox analysis that adjusted for duration and wound area. Kaplan-Meier analysis was used to determine median time to complete wound closure (CWC). RESULTS: The cohort comprised 218 patients, and these were split equally between the two treatment groups with 109 patients in each. A Cox analysis showed that the estimated frequency of wound closure for the dACM plus SoC group was statistically superior to the SoC alone group at week 4 (12% versus 8%), week 6 (22% versus 11%), week 8 (31% versus 21%), week 10 (42% versus 27%) and week 12 (50% versus 35%), respectively (p=0.04). The computed hazard ratio (1.48 (confidence interval: 0.95, 2.29) showed a 48% greater probability of wound closure in favour of the dACM group. Median time to wound closure for dACM-treated ulcers was 84 days compared to 'not achieved' in the SoC-treated group (i.e., ≥50% of SoC-treated DFUs failed to heal by week 12; p=0.04). CONCLUSION: In an adequately powered DFU RCT, dACM increased the frequency, decreased the median time, and improved the probability of CWC when compared with SoC alone. dACM demonstrated beneficial effects in DFUs in a complex patient population. DECLARATION OF INTEREST: This study was funded by Organogenesis Inc., US. JC serves as a consultant and speaker for Organogenesis. RDD serves as a speaker for Organogenesis. OMA and MLS serve as consultants for Organogenesis. The authors have no other conflicts of interest to declare.


Assuntos
Âmnio , Córion , Pé Diabético , Padrão de Cuidado , Cicatrização , Humanos , Pé Diabético/terapia , Feminino , Âmnio/transplante , Masculino , Córion/transplante , Pessoa de Meia-Idade , Estudos Prospectivos , Idoso , Resultado do Tratamento , Adulto , Curativos Biológicos
8.
J Wound Care ; 33(Sup3): S24-S38, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38457290

RESUMO

OBJECTIVE: To evaluate the cost-effectiveness of dehydrated human amnion/chorion membrane (DHACM) in Medicare enrolees who developed a venous leg ulcer (VLU). METHOD: This economic evaluation used a four-state Markov model to simulate the disease progression of VLUs for patients receiving advanced treatment (AT) with DHACM or no advanced treatment (NAT) over a three-year time horizon from a US Medicare perspective. DHACM treatments were assessed when following parameters for use (FPFU), whereby applications were initiated 30-45 days after the initial VLU diagnosis claim, and reapplications occurred on a weekly to biweekly basis until completion of the treatment episode. The cohort was modelled on the claims of 530,220 Medicare enrolees who developed a VLU between 2015-2019. Direct medical costs, quality-adjusted life years (QALYs), and the net monetary benefit (NMB) at a willingness-to-pay threshold of $100,000/QALY were applied. Univariate and probabilistic sensitivity analyses (PSA) were performed to test the uncertainty of model results. RESULTS: DHACM applied FPFU dominated NAT, yielding a lower per-patient cost of $170 and an increase of 0.010 QALYs over three years. The resulting NMB was $1178 per patient in favour of DHACM FPFU over the same time horizon. The rate of VLU recurrence had a notable impact on model uncertainty. In the PSA, DHACM FPFU was cost-effective in 63.01% of simulations at the $100,000/QALY threshold. CONCLUSION: In this analysis, DHACM FPFU was the dominant strategy compared to NAT, as it was cost-saving and generated a greater number of QALYs over three years from the US Medicare perspective. A companion VLU Medicare outcomes analysis revealed that patients who received AT with a cellular, acellular and matrix-like product (CAMP) compared to patients who received NAT had the best outcomes. Given the added clinical benefits to patients at lower cost, providers should recommend DHACM FPFU to patients with VLU who qualify. Decision-makers for public insurers (e.g., Medicare and Medicaid) and commercial payers should establish preferential formulary placement for reimbursement of DHACM to reduce budget impact and improve the long-term health of their patient populations dealing with these chronic wounds. DECLARATION OF INTEREST: Support for this analysis was provided by MiMedx Group, Inc., US. JLD, and RAF are employees of MiMedx Group, Inc. WHT, BH, PS, BGC and WVP were consultants to MiMedx Group, Inc. VD, AO, MRK, JAN, NW and GAM served on the MiMedx Group, Inc. Advisory Board. MRK and JAN served on a speaker's bureau. WVP declares personal fees and equity holdings from Stage Analytics, US.


Assuntos
Análise de Custo-Efetividade , Úlcera Varicosa , Idoso , Humanos , Estados Unidos , Âmnio , Cicatrização , Córion , Medicare , Úlcera Varicosa/terapia , Análise Custo-Benefício
9.
Cell Tissue Bank ; 25(1): 349-356, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36840842

RESUMO

The purpose of the present study was to process and assess the effect of hydrated amnion chorion membrane and dehydrated amnion chorion membrane on proliferation of periodontal ligament (PDL) fibroblast cells. The amnion chorion membrane (ACM) from placenta of 18 systemically healthy patients was obtained from the Department of Obstetrics and Gynaecology. They were processed as hydrated and dehydrated based on different processing methods. The Periodontal ligament cells were obtained from periodontal ligament of freshly extracted premolars of systemically healthy patients, due to orthodontic reasons. The PDL cells were further cultured in laboratory and were exposed to hydrated and dehydrated amnion chorion membrane. The MTT assay was performed to assess the proliferation of PDL fibroblast cells after 24 and 48 h. The hydrated and dehydrated amnion chorion membrane showed proliferation of PDL fibroblasts after 24 and 48 h. The proliferation of PDL fibroblasts in hydrated (p = 0.043) and dehydrated (p = 0.050) amnion chorion membrane was statistically significant at the end of 24 and 48 h respectively. On inter-group comparison dehydrated ACM showed significant proliferation of PDL fibroblasts after 24 (p=0.014) and 48 h (p=0.019). Within the limits of the present study, it can be concluded: both hydrated and dehydrated amnion chorion membrane showed proliferationof PDL fibroblast cells. However, dehydrated ACM showed significant proliferation of PDL fibroblasts.


Assuntos
Âmnio , Cicatrização , Gravidez , Feminino , Humanos , Ligamento Periodontal , Fibroblastos , Córion , Proliferação de Células
10.
Int Wound J ; 21(10): e70096, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39402007

RESUMO

Diabetic foot ulcers (DFUs) are a severe complication for diabetic patients, significantly impacting patient quality of life and healthcare system efficiency. Traditional standard of care (SOC) treatments are inadequate for many patients, necessitating the use of advanced wound care products, such as human placental membranes. We studied a real-world population of large, hard-to-heal and complicated wounds, otherwise under-studied in the wound care literature. To this end, we conducted a retrospective cohort analysis to compare the effectiveness of a human placental amnion/chorion membrane product using retention-based processing (RE-AC) and SOC in managing chronic DFUs. During the study period of September 2021 through April 2024, we collected retrospective observational data from electronic health records of 21 patients treated with RE-AC at three outpatient wound care centres. Additionally, 21 control SOC patients were matched from a wound registry using Coarsened Exact Matching. Patients were categorized into two cohorts based on whether they received RE-AC or SOC. Key metrics included wound size progression and wound closure. The analysis employed Bayesian regression and Hurdle Gamma Analysis of Covariance models. Despite their rather large size (average of 13.8 cm2), our results indicated that RE-AC achieved almost a 10% higher expected wound closure rate compared to SOC at 12 weeks (8.53% [credible interval: 5.60%-10.7%]). Further, for wounds that did not close, RE-AC resulted in a 93.6% (credible interval: 147.7%-41.6) improvement in expected Percent Area Reduction over the SOC group at 12 weeks. We noted that on average, SOC wounds stalled or grew larger. In terms of a risk ratio comparing the study group with SOC, we found a 52% benefit in the RE-AC group (RR = 1.52). The findings suggest that even with larger DFUs, R-AC is superior to SOC for wound closure and expected Percent Area Reduction by 12 weeks. This benefit likely leads to reduced treatment costs, optimized resource utilization and improved outcomes in the DFU patient population; ultimately resulting in improved patient care.


Assuntos
Pé Diabético , Padrão de Cuidado , Cicatrização , Humanos , Pé Diabético/terapia , Estudos Retrospectivos , Feminino , Pessoa de Meia-Idade , Masculino , Idoso , Placenta , Gravidez , Âmnio/transplante , Córion , Adulto , Estudos de Coortes
11.
Int Wound J ; 21(1): e14356, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37661177

RESUMO

We investigated the healing effect of a new dehydrated amnion/chorion membrane with a spongy layer over a 30-month period in 32 patients with 53 chronic non-healing wounds of different aetiologies. Wounds with <40% surface reduction after 4 weeks of best wound treatment underwent weekly allograft application by a certified wound specialist based on national guidelines and a standardised protocol until complete healing or definite treatment interruption. The main outcome measure was the percentage of wound surface reduction from baseline calculated using digital planimetry follow-up photographs. Overall, 38 (71.7%) wounds presented a favourable outcome (70%-100% area reduction), with 35 (66%) completely healing over a median time of 77 days (range 29-350 days). Favourable outcomes were observed in 75% of traumatic wounds, surgical wounds, venous leg ulcers and pressure injuries, as well as in 50% of ischaemic wounds. Wounds being present <12 months were significantly more likely to have a favourable outcome than more long-standing wounds (χ2 = 7.799; p = 0.005; OR = 3.378; 95% CI, 1.410-8.092). Thus, treatment with dehydrated amnion/chorion membrane with a spongy layer improves the outcome of non-healing wounds of different aetiologies and, therefore, has to be considered early in the management of refractory wounds.


Assuntos
Âmnio , Córion , Humanos , Aloenxertos/transplante , Âmnio/transplante , Resultado do Tratamento , Transplante Homólogo/métodos , Córion/transplante
12.
J Contemp Dent Pract ; 25(2): 160-167, 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38514414

RESUMO

AIM: Amnion and chorion membranes possess unique inherited biological properties that enhance wound healing and may accelerate periodontal regeneration. The present study aims to evaluate and compare the efficacy of amnion and chorion membranes in the treatment of furcation defects. MATERIALS AND METHODS: A total of 20 patients were selected and were randomly allocated to group I and group II with 10 subjects in each group. Amnion and chorion membranes are placental-derived membranes that accelerate regeneration by having natural growth factors with their antimicrobial and inflammation reduction properties. Group I was treated using bone grafting with decalcified freeze-dried bone allograft (DFDBA) and placement of amnion as a membrane for guided tissue regeneration (GTR) whereas group II was treated using bone grafting with DFDBA and placement of chorion as a membrane for GTR. The patients were followed for clinical and radiographic parameters and were evaluated between 3 and 6 months after surgery. RESULT: In intragroup comparison, a significant difference was evident in both the groups for all the clinical and radiographic parameters within the groups. (p = 0.01) This means both amnion and chorion membranes showed statistically significant regenerative efficacy. In intergroup comparison, the results show that all the clinical parameters and radiographic parameters show no significant difference between the groups. CONCLUSION: The amnion and chorion membranes had similar regenerative efficacy in combination with DFDBA in patients with buccal degree II furcation defects in mandibular molars. CLINICAL SIGNIFICANCE: The amnion and chorion membranes have shown significant improvement in clinical and radiographic parameters when used for the treatment of buccal degree II furcation defects in mandibular molars. How to cite this article: Mallapragda S, Gupta R, Gupta S, et al. Evaluation of Regenerative Efficacy of Amnion and Chorion Membrane in Treatment of Mandibular Molar Furcation Defects: A Clinico-radiographic Study. J Contemp Dent Pract 2024;25(2):160-167.


Assuntos
Defeitos da Furca , Gravidez , Humanos , Feminino , Defeitos da Furca/cirurgia , Âmnio/transplante , Regeneração Tecidual Guiada Periodontal/métodos , Placenta/cirurgia , Dente Molar/cirurgia , Transplante Ósseo/métodos , Córion/cirurgia , Membranas Artificiais
13.
Development ; 147(4)2020 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-32054660

RESUMO

La-related protein 6 (Larp6) is a conserved RNA-binding protein found across eukaryotes that has been suggested to regulate collagen biogenesis, muscle development, ciliogenesis, and various aspects of cell proliferation and migration. Zebrafish have two Larp6 family genes: larp6a and larp6b Viable and fertile single and double homozygous larp6a and larp6b zygotic mutants revealed no defects in muscle structure, and were indistinguishable from heterozygous or wild-type siblings. However, larp6a mutant females produced eggs with chorions that failed to elevate fully and were fragile. Eggs from larp6b single mutant females showed minor chorion defects, but chorions from eggs laid by larp6a;larp6b double mutant females were more defective than those from larp6a single mutants. Electron microscopy revealed defective chorionogenesis during oocyte development. Despite this, maternal zygotic single and double mutants were viable and fertile. Mass spectrometry analysis provided a description of chorion protein composition and revealed significant reductions in a subset of zona pellucida and lectin-type proteins between wild-type and mutant chorions that paralleled the severity of the phenotype. We conclude that Larp6 proteins are required for normal oocyte development, chorion formation and egg activation.


Assuntos
Autoantígenos/genética , Autoantígenos/fisiologia , Córion/fisiologia , Oócitos/fisiologia , Ribonucleoproteínas/genética , Ribonucleoproteínas/fisiologia , Animais , Movimento Celular , Proliferação de Células , Colágeno/fisiologia , Proteínas do Ovo/fisiologia , Feminino , Edição de Genes , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Genoma , Genótipo , Heterozigoto , Homozigoto , Lectinas/fisiologia , Masculino , Mutação , Oócitos/citologia , Oogênese/fisiologia , Fenótipo , Peixe-Zebra , Zona Pelúcida/fisiologia , Antígeno SS-B
14.
Insect Mol Biol ; 32(5): 484-509, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37158315

RESUMO

Chorion-i.e., the eggshell-is formed during the late stage of oogenesis by follicular epithelium in the ovary. Although the endocrine signal(s) driving choriogenesis remain unclear in mosquitoes, this process in other insects has been suspected to involve the mediation of prostaglandins (PGs). This study tested the role of PG in the choriogenesis of the Asian tiger mosquito, Aedes albopictus, and its influence on controlling the expressions of genes associated with chorion formation by a transcriptome analysis. An immunofluorescence assay showed that PGE2 is localised in follicular epithelium. With the treatment of aspirin, an inhibitor of PG biosynthesis, at mid oogenesis, the PGE2 signal disappeared in the follicular epithelium led to significantly inhibited chorion formation along with a malformed eggshell. Ovary transcriptomes were assessed by RNASeq at the mid and late ovarian developmental stages. Differentially expressed genes (DEGs) exhibiting more than twofold changes in expression levels included 297 genes at mid stage and 500 genes at late stage. These DEGs at these two developmental stages commonly included genes associated with egg and chorion proteins of Ae. albopictus. Most chorion-associated genes were clustered in the 168 Mb region on a chromosome and exhibited significantly induced expressions at both ovarian developmental stages. The inhibition of PG biosynthesis significantly suppressed the expression of the chorion-associated genes while the addition of PGE2 rescued the gene expressions and led to recovery of choriogenesis. These results suggest that PGE2 mediates the choriogenesis of Ae. albopictus.


Assuntos
Aedes , Feminino , Animais , Aedes/metabolismo , Oogênese , Ovário , Prostaglandinas/metabolismo , Córion , Mosquitos Vetores
15.
Wound Repair Regen ; 31(4): 475-488, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37209062

RESUMO

Recent evidence suggests that protecting human amnion and chorion matrices (HACM) during processing enhances the performance of HACM for wound repair and tissue regeneration. We utilised a diabetic (db/db) delayed wound healing mouse model. Treatment of db/db full-thickness excisional wounds with HACM, processed with a polyampholyte preservative accentuated the proliferative phase of wound healing that decreased the time necessary to heal wounds. Polyampholyte protection improved the preservation of growth factors and cytokines during room temperature storage following E-beam sterilisation and improved its function in wound healing applications. Our findings indicate protected HACM tissue up-regulated MIP2, NF-kB, TNF-α, KI-67, and Arg1 (0.6-fold to 1.5-fold) but those changes were not statistically significant. Immunofluorescent assessment identifying cell activity illustrated an induction of the proliferative phase of wound healing and a switch from an inflammatory macrophage phenotype (M1) to a pro-regenerative macrophage phenotype (M2a). Genomic profiling of 282 genes was performed using Nanostring from co-cultures of human macrophages and fibroblasts. The polyampholyte + HACM-treated group, compared with the HACM or polyampholyte alone groups, had a statistically significant up-regulation (32-368 fold) of 12 genes primarily involved in macrophage plasticity including CLC7, CD209, CD36, HSD11B1, ICAM1, IL1RN, IL3RA, ITGAX, LSP1, and PLXDC2 (adj. p-value < 0.05). The polyampholyte alone group demonstrated statistically significant down-regulation of four genes ADRA2, COL7A1, CSF3, and PTGS2 (adj. p < 0.05). The HACM alone group up-regulated four genes ATG14, CXCL11, DNMT3A, and THBD, but the results were not statistically significant. Biomechanical measurements indicated that wounds treated with polyampholyte-protected HACM had more tensile integrity compared with wounds treated with HACM alone. These findings indicate that better protection of HACM during processing stabilises the HACM matrix, which may lead to improved wound healing outcomes.


Assuntos
Âmnio , Diabetes Mellitus , Camundongos , Animais , Humanos , Técnicas de Cocultura , Cicatrização/fisiologia , Córion , Modelos Animais de Doenças , Colágeno Tipo VII
16.
Ecotoxicol Environ Saf ; 264: 115446, 2023 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-37688866

RESUMO

Concerns over the spread of non-native species in aquatic environments have led to the need for effective methods to prevent and control their spread while protecting native species. This study investigated the potential of yeast vacuolar enzymes as a natural hatching inhibitor for controlling aquatic organisms. Hatching experiments with Daphnia magna eggs demonstrated that exposure to yeast vacuole enzymes inhibited hatching in a concentration-dependent manner, suggesting their potential as an effective inhibitor of egg hatching in aquatic organisms. Interestingly, the protease used for comparative purposes did not inhibit hatching, but instead increased the mortality of hatched D. magna. Additionally, chorionic changes were observed in non-hatched D. magna eggs and zebrafish eggs exposed to yeast vacuole enzymes, suggesting that the enzyme can alter the chorion and interfere with hatching. These findings suggest that yeast vacuolar enzymes may be a promising and natural management tool for controlling the spread of harmful aquatic organisms, and further research is warranted to explore their potential for species-specific control.


Assuntos
Saccharomyces cerevisiae , Peixe-Zebra , Animais , Daphnia , Organismos Aquáticos , Vacúolos
17.
J Wound Care ; 32(10): 634-640, 2023 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-37830838

RESUMO

Decompressive craniectomies (DCs) are routinely performed neurosurgical procedures to emergently treat increased intracranial pressure secondary to multiple aetiologies, such as subdural haematoma, epidural haematoma, or malignant oedema in the setting of acute infarction. The DC procedure typically induces epidural fibrosis post-cranial resection, resulting in adherence of the dura to both the brain internally and skin flap externally. This becomes especially problematic in the setting of skull flap replacement for cranioplasty as adherences can lead to bridging vein tear, damage to the underlying brain cortex, and other postoperative complications. Dural adjuvants, which can contribute to decreased rate of adherence formation, can thereby reduce both postoperative cranioplasty complications and operative duration. Dehydrated human amnion/chorion membrane (DHACM) allografts (AMNIOFIX, MIMEDX Group Inc., US) have been shown to reduce the rate of dural scar tissue formation in re-exploration of posterior lumbar interbody fusion operations which require entry into the epidural space. The purpose of this study was to evaluate whether or not the use of DHACM in the setting of emergent craniectomies decreased the rate of dural adhesion formation and subsequent cranioplasty complications. Patients (n=7) who underwent emergent craniectomy and intraoperative placement of DHACM were evaluated during replacement of either an autologous skull cap or a custom-made implant, at which point the degree of adhesions was qualitatively assessed. Placement of DHACM below and on top of the dura resulted in negligible adhesion being found during the defect exposure, and there were no intraoperative complications during cranioplasties. Reported estimated blood loss across the seven patients averaged 64.2ml, total operative time averaged 79.2 minutes, and time dedicated to exposing defect for bone flap placement was <3 minutes.


Assuntos
Âmnio , Procedimentos de Cirurgia Plástica , Humanos , Âmnio/transplante , Craniotomia/efeitos adversos , Retalhos Cirúrgicos , Aderências Teciduais/cirurgia , Aderências Teciduais/etiologia , Complicações Pós-Operatórias/etiologia , Córion/transplante
18.
Pediatr Surg Int ; 40(1): 7, 2023 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-37999778

RESUMO

BACKGROUND: Patch repair of congenital diaphragmatic hernia (CDH) using Gore-Tex® is associated with infection, adhesions, hernia recurrence, long-term musculoskeletal sequels and poor tissue regeneration. To overcome these limitations, the performance of two novel biodegradable membranes was tested to repair CDH in a growing pig model. METHODS: Twelve male pigs were randomly assigned to 3 different groups of 4 animals each, determined by the type of patch used during thoracoscopic diaphragmatic hernia repair (Gore-Tex®, polycaprolactone electrospun membrane-PCLem, and decellularized human chorion membrane-dHCM). After 7 weeks, all animals were euthanized, followed by necropsy for diaphragmatic evaluation and histological analysis. RESULTS: Thoracoscopic defect creation and diaphragmatic repair were performed without any technical difficulty in all groups. However, hernia recurrence rate was 0% in Gore-Tex®, 50% in PCLem and 100% in dHCM groups. At euthanasia, Gore-Tex® patches appeared virtually unchanged and covered with a fibrotic capsule, while PCLem and dHCM patches were replaced by either floppy connective tissue or vascularized and floppy regenerated membranous tissue, respectively. CONCLUSION: Gore-Tex® was associated with a higher survival rate and lower recurrence. Nevertheless, the proposed biodegradable membranes were associated with better tissue integration when compared with Gore-Tex®.


Assuntos
Hérnias Diafragmáticas Congênitas , Politetrafluoretileno , Animais , Masculino , Diafragma , Hérnias Diafragmáticas Congênitas/cirurgia , Herniorrafia , Suínos
19.
Int J Mol Sci ; 24(21)2023 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-37958770

RESUMO

Placental membranes have been widely studied and used clinically for wound care applications, but there is limited published information on the benefits of using the chorion membrane. The chorion membrane represents a promising source of placental-derived tissue to support wound healing, with its native composition of extracellular matrix (ECM) proteins and key regulatory proteins. This study examined the impact of hypothermic storage on the structure of chorion membrane, ECM content, and response to degradation in vitro. Hypothermically stored chorion membrane (HSCM) was further characterized for its proteomic content, and for its functionality as a scaffold for cell attachment and proliferation in vitro. HSCM retained the native ECM structure, composition, and integrity of native unprocessed chorion membrane and showed no differences in response to degradation in an in vitro wound model. HSCM retained key regulatory proteins previously shown to be present in placental membranes and promoted the attachment and proliferation of fibroblasts in vitro. These data support the fact that hypothermic storage does not significantly impact the structure and characteristics of the chorion membrane compared to unprocessed tissue or its functionality as a scaffold to support tissue growth.


Assuntos
Placenta , Proteômica , Humanos , Feminino , Gravidez , Âmnio , Proliferação de Células/fisiologia , Cicatrização/fisiologia , Córion , Proteínas da Matriz Extracelular/análise
20.
Wiad Lek ; 76(8): 1797-1803, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37740973

RESUMO

OBJECTIVE: The aim: To examine the morphology regarding the term placentas in pregnancies with threatened abortion with bleeding in the first trimester. PATIENTS AND METHODS: Materials and methods: 118 term placentas were selected, of which 40 placentas with the physiological course of pregnancy. 78 placentas were from women with threatened abortion with bleeding in the first trimester, of which 37 patients received hormonal therapy (group I), 41 women were prescribed symptomatic therapy (group II). Placentas were investigatedaccording to the protocol, which includes organometric, macroscopic and microscopic studies. RESULTS: Results: In the placentas of group II is a significant increase of the area of terminal villi compared due to the stroma against the background of a deficit of fetal capillaries. In group I have revealed that the specific weight of the vascular bed of the terminal villi was 1.5 times higher compared to the control (Р=0.031) and 2.7 times higher than the group II (Р=0.022) and dominates the share of the stroma. The weight of the epithelium of the terminal villi in all groups is approximately the same (Р=0.042), but the ratio of the epithelium to the stroma is higher in the group I (0.63) than in the control (0.43). CONCLUSION: Conclusions: In women with a pathological course of the first trimester of pregnancy the compensatory mechanisms in full-term placentas are morphologically represented by an increase in the number of terminal villi, syncytio-capillary membranes, intensification of angiogenesis. In the placentas of women who received hormonal therapy adaptive reactions are most effective and able to compensate for the gestational immaturity of the chorion.


Assuntos
Ameaça de Aborto , Gravidez , Humanos , Feminino , Primeiro Trimestre da Gravidez , Hemorragia , Duodeno , Placenta
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